Methods for storing cysteamine formulations and related methods of treatment

ABSTRACT

Methods of storing and methods of stabilizing pharmaceutical compositions comprising cysteamine, or a pharmaceutically acceptable salt thereof, are provided. Methods of distributing pharmaceutical compositions comprising cysteamine, or a pharmaceutically acceptable salt thereof, and methods of treating cystinosis also are provided.

BACKGROUND

Cystinosis is a rare, autosomal recessive disease caused byintra-lysosomal accumulation of the amino acid cysteine within varioustissues, including the spleen, liver, lymph nodes, kidney, bone marrow,and eyes. Nephropathic cystinosis is associated with kidney failure thatnecessitates kidney transplantation. A specific treatment fornephropathic cystinosis is the sulfhydryl agent cysteamine. Cysteaminehas been shown to lower intracellular cystine levels, thereby reducingthe rate of progress of kidney failure in children.

Cysteamine, and pharmaceutically acceptable salts thereof, may also beadministered for the treatment of other metabolic and neurodegenerativediseases, including non-alcoholic fatty liver disease (NAFLD),non-alcoholic steatohepatitis (NASH), Huntington's disease, Parkinson'sdisease, Rett Syndrome, cystic fibrosis, and others; used as freeradical and radioprotectants; used as hepato-protectant agents.

Enterically-coated cysteamine compositions for increasing delivery ofcysteamine to the small intestine and resulting in less frequent dosingcompared to non-enteric-coated cysteamine have been described. See,e.g., WO 2014/204881, WO 2007/089670, and U.S. Pat. Nos. 8,026,284,9,198,882, 9,192,590, 9,173,851, and 9,233,077.

In some cysteamine pharmaceutical compositions, cysteamine is notchemically stable and degrades into several impurities over time.

SUMMARY OF THE DISCLOSURE

The present disclosure provides methods of storing pharmaceuticalcompositions comprising cysteamine, or a pharmaceutically acceptablesalt thereof, wherein the pharmaceutical composition is stored atrefrigerated temperatures (e.g., 2° C.-8° C.) up to 24 months, orlonger, and compositions that have been stored in such a manner thathave fewer impurities than when stored at 25° C., and methods for usingthe same. In various embodiments, the disclosure provides a method ofstoring a pharmaceutical composition, comprising storing thepharmaceutical composition at a temperature of between about 2° C. andabout 8° C., wherein the pharmaceutical composition comprisescysteamine, or a pharmaceutically acceptable salt thereof.

In one embodiment, the disclosure provides a method of stabilizing apharmaceutical composition, comprising storing the pharmaceuticalcomposition at a temperature of between about 2° C. and about 8° C.,wherein the pharmaceutical composition comprises cysteamine, or apharmaceutically acceptable salt thereof.

Also provided is a method of distributing a pharmaceutical composition,comprising storing the pharmaceutical composition at a temperature ofbetween about 2° C. and about 8° C. prior to dispensing to a health careprovider or a patient, wherein the pharmaceutical composition comprisescysteamine, or a pharmaceutically acceptable salt thereof.

Further contemplated is a method of treating a disease or disorder,comprising administering a pharmaceutical composition comprisingcysteamine, or a pharmaceutically acceptable salt thereof to a subjectin need thereof, wherein the pharmaceutical composition has been storedat a temperature of between about 2° C. and about 8° C. prior toadministration. Exemplary diseases or disorders contemplated hereininclude, but are not limited to, cystinosis, fatty liver disease, athrombotic disease, an MECP-2 related disorder, an inheritedmitochondrial disease, a neurological disease or disorder, inflammationand cancer. Additional indications contemplated are set out in theDetailed Description.

In various embodiments of the methods, the pharmaceutical compositionfurther comprises one or more materials that provide increased deliveryof cysteamine to the small intestine. In one embodiment, a material thatprovides increased delivery of cysteamine to the small intestinecomprises an enteric coating. Exemplary enteric coatings contemplatedcomprise a coating selected from the group consisting of polymerizedgelatin, shellac, methacrylic acid copolymer type CNF, cellulosebutyrate phthalate, cellulose hydrogen phthalate, cellulose proprionatephthalate, polyvinyl acetate phthalate (PVAP), cellulose acetatephthalate (CAP), cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate, hydroxypropyl methylcellulose acetate,dioxypropyl methylcellulose succinate, carboxymethyl ethylcellulose(CMEC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), andacrylic acid polymers and copolymers formed from methyl acrylate, ethylacrylate, methyl methacrylate, and/or ethyl methacrylate with copolymersof acrylic and methacrylic acid esters. In one embodiment, the entericcoating comprises poly(methacrylic acid co-ethyl acrylate) 1:1 (EudragitL 30-D-55).

In various embodiments of the methods, the pharmaceutical compositioncomprises a pharmaceutically acceptable salt of cysteamine, and thepharmaceutically acceptable salt of cysteamine is cysteamine bitartrate.

It is also contemplated that the pharmaceutical composition comprises asolid composition. In one embodiment, the pharmaceutical compositioncomprises a unit dose of about 25 mg cysteamine or of about 75 mgcystemine.

In various embodiments, the pharmaceutical composition comprises entericcoated beads. In various embodiments, the pharmaceutical composition isa pharmaceutical dosage form that includes a plurality of cysteaminebeads, the beads including a core particle including cysteamine or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable excipient, and an enteric membrane surrounding the coreparticle, wherein the plurality of beads is characterized by adistribution of particle sizes.

In one embodiment, the particle sizes of the beads are in a range ofabout 0.7 mm to about 2.5 mm, or about 0.7 mm to about 2.8 mm, or about0.8 mm to about 1.7 mm. For example, the target bead size can be up to2.5 mm with no more than 10 percent variation over this size, to amaximum size of 2.8 mm. In one embodiment, the enteric membrane on thebeads is present in an amount in a range of about 20% to 40%, or about20% to about 25%, or about 25% to about 35% as measured by the weightgain compared to the uncoated particle cores, or in a range of about 25%to about 31% weight gain, or about 27% to about 31% weight gain, orabout 28.5% to about 31% weight gain, based on the weight of theuncoated particle cores.

In various embodiments, the bead formulation comprises a plurality ofcysteamine beads, the beads comprising a core particle comprising,cysteamine or a pharmaceutically acceptable salt thereof, such ascysteamine bitartrate, a filler, a binder and an enteric membranesurrounding the core, wherein the plurality of beads is characterized bya distribution of particle sizes in a range of about 0.7 mm to about 2.8mm; wherein the enteric membrane is present in an amount in a range ofabout 20% to about 40% based on the weight of the bead core particles;wherein the formulation is a delayed release formulation having anenteric membrane that begins to dissolve within a pH range of about 4.5to about 6.5, and wherein the beads are disposed in a capsule shell.

In further embodiments of the methods above, the pharmaceuticalcomposition is stored at a temperature of between about 2° C. and about8° C. for up to 1 month, between about 1 month and about 6 months,between about 6 months and about 12 months, between about 12 month andabout 15 months, between about 15 months and about 18 months, betweenabout 18 months and about 21 months, between about 21 months and about24 months, between about 24 months and about 36 months, or between about36 months and about 39 months. In various embodiments, thepharmaceutical composition is stored at a temperature of between about2° C. and about 8° C. for 1 month, 6 months, 12 months, 15 months, 18months, 21 months, 24 months, 36 months, or 39 months.

The methods herein may further comprise storing the pharmaceuticalcomposition at a temperature of between about 20° C. and about 25° C.and a relative humidity of 60% for up to 4 months.

In various embodiments, the pharmaceutical composition is stable through12, 15, 18, 21 or 24 months at 2° C.-8° C. storage. In variousembodiments, the pharmaceutical composition is stable through 24 monthsat 2° C.-8° C. storage. In various embodiments, the pharmaceuticalcomposition is stable after storage at a temperature of 2° C.-8° C. forup to 15 months followed by excursions of up to 3 months at 25° C./60%RH, 30° C./65% RH, or 30° C./75% RH (18 months of total storage time).

In various embodiments of the methods, the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, is less that0.5%. In some embodiments, the amount of 2-hydroxythiomorpholine presentin the pharmaceutical composition, relative to the amount of cysteamine,or a pharmaceutically acceptable salt thereof, present in thepharmaceutical composition, is less than or equal to the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, after storageat a temperature of between about 20° C. and about 25° C. and a relativehumidity of 60% for the same duration.

In various embodiments of the methods above, the amount of cystaminepresent in the pharmaceutical composition, relative to the amount ofcysteamine, or a pharmaceutically acceptable salt thereof, present inthe pharmaceutical composition, is less that 4%. In certain embodiments,the amount of cystamine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, is less than orequal to the amount of cystamine present in the pharmaceuticalcomposition, relative to the amount of cysteamine, or a pharmaceuticallyacceptable salt thereof, present in the pharmaceutical composition,after storage at a temperature of between about 20° C. and about 25° C.and a relative humidity of 60% for the same duration.

It is contemplated that the amount of cystamine tartrate amide presentin the pharmaceutical composition, relative to the amount of cysteamine,or a pharmaceutically acceptable salt thereof, present in thepharmaceutical composition, is less than 0.5%. In various embodiments,the amount of cystamine tartrate amide present in the pharmaceuticalcomposition, relative to the amount of cysteamine, or a pharmaceuticallyacceptable salt thereof, present in the pharmaceutical composition, isless than or equal to the amount of cystamine tartrate amide present inthe pharmaceutical composition, relative to the amount of cysteamine, ora pharmaceutically acceptable salt thereof, present in thepharmaceutical composition, after storage at a temperature of betweenabout 20° C. and about 25° C. and a relative humidity of 60% for thesame duration.

The methods also provide that the amount of 2-hydroxymethylthiazolidinepresent in the pharmaceutical composition, relative to the amount ofcysteamine, or a pharmaceutically acceptable salt thereof, present inthe pharmaceutical composition is less than 0.05%. In variousembodiments, the amount of 2-hydroxymethylthiazolidine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, is less than or equal to the amount of2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, after storageat a temperature of between about 20° C. and about 25° C. and a relativehumidity of 60% for the same duration.

In various embodiments, the total amount of 2-hydroxymethylthiazolidine,cystamine, cystamine tartrate amide, and 2-hydroxymethylthiazolidinepresent in the pharmaceutical composition, relative to the amount ifcysteamine, or a pharmaceutically acceptable salt thereof, present inthe pharmaceutical composition, is less than or equal to the totalamount of 2-hydroxymethylthiazolidine, cystamine, cystamine tartrateamide, and 2-hydroxymethylthiazolidine present in the pharmaceuticalacceptable salt thereof, present in the pharmaceutical composition,after storage at a temperature of between about 20° C. and about 25° C.and relative humidity of 60% for the same duration.

In another aspect, the present disclosure provides methods of treating adisease or disorder by administering to a subject in need thereof apharmaceutical composition comprising cysteamine, or a pharmaceuticallyacceptable salt thereof, wherein the pharmaceutical composition has beenstored at a temperature of between about 2° C. and about 8° C. prior toadministration. Disease or disorders contemplated herein are describedin more detail in the Detailed Description.

Also provided is a method of treating cystinosis, comprisingadministering a pharmaceutical composition comprising cysteamine, or apharmaceutically acceptable salt thereof, wherein the pharmaceuticalcomposition has been stored at a temperature of between about 2° C. andabout 8° C. prior to administration. In various embodiments, thecystinosis is nephropathic cystinosis.

In various embodiments, the pharmaceutical composition is formulated toprovide white blood cell cystine suppression with a 12 hour level below1 nmol 1/2 cystine/mg protein.

In various embodiments, each dose of cysteamine is about 0.5-1.0 g/m2body surface area. In various embodiments, the total daily dose ofcysteamine is about 1.3 g/m2 body surface area or less. In variousembodiments, the composition is administered less than four times daily,e.g., one, two or three times a day. In various embodiments, thecomposition is administered twice daily, or every 12 hours.

In various embodiments, the composition increases delivery to theproximal small intestine, the mid-small intestine, the duodenum, thejejunum or the mid-ileum.

In various embodiments, the composition is in the form of a tablet or acapsule.

In various embodiments, the cysteamine salt is cysteamine bitartrate.

DESCRIPTION OF THE DRAWINGS

FIG. 1A shows a comparison of cold storage (2° C. to 8° C.) to roomtemperature storage (25° C./60% RH) for the amount of cysteaminebitartrate (relative the label claim; expressed as a percent) in samplesof 25-mg PROCYSBI® (“Assay”). Open circles and squares (∘, □) withdotted lines represent 25° C./60% RH data for lots A and B,respectively. Closed circles and squares (●, ▪) with solid linesrepresent 2° C.-8° C. data for lots A and B, respectively.

FIG. 1B shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for acid stage dissolution tests ofsamples of 25-mg PROCYSBI® (relative the label claim; expressed as apercent). Open circles and squares (∘, □) with dotted lines represent25° C./60% RH data for lots A and B, respectively. Closed circles andsquares (●, ▪) with solid lines represent 2° C.-8° C. data for lots Aand B, respectively.

FIG. 1C shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for buffer stage dissolution testsof samples of 25-mg PROCYSBI® (relative the label claim; expressed as apercent). Open circles and squares (∘, □) with dotted lines represent25° C./60% RH data for lots A and B, respectively. Closed circles andsquares (●, ▪) with solid lines represent 2° C.-8° C. data for lots Aand B, respectively.

FIG. 1D shows comparisons of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for cystamine (relative tocysteamine bitartrate; expressed as a percentage) in samples of 25-mgPROCYSBI®. Open circles and squares (∘, □) with dotted lines represent25° C./60% RH data for lots A and B, respectively. Closed circles andsquares (●, ▪) with solid lines represent 2° C.-8° C. data for lots Aand B, respectively.

FIG. 1E shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for 2-hydroxythiomorpholine(relative to cysteamine bitartrate; expressed as a percentage) insamples of 25-mg PROCYSBI®. Open circles and squares (∘, □) with dottedlines represent 25° C./60% RH data for lots A and B, respectively.Closed circles and squares (●, ▪) with solid lines represent 2° C.-8° C.data for lots A and B, respectively.

FIG. 1F shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for cystamine tartrate amide(relative to cysteamine bitartrate; expressed as a percentage) insamples of 25-mg PROCYSBI®. Open circles and squares (∘, □) with dottedlines represent 25° C./60% RH data for lots A and B, respectively.Closed circles and squares (●, ▪) with solid lines represent 2° C.-8° C.data for lots A and B, respectively.

FIG. 1G shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for Peak 6 (relative to cysteaminebitartrate; expressed as a percentage) in samples of 25-mg PROCYSBI®.Open circles and squares (∘, □) with dotted lines represent 25° C./60%RH data for lots A and B, respectively. Closed circles and squares (●,▪) with solid lines represent 2° C.-8° C. data for lots A and B,respectively.

FIG. 2A shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for cystamine (relative tocysteamine bitartrate; expressed as a percentage) in samples of 25-mg or75-mg PROCYSBI®. Open symbols with dotted lines represent 25° C./60% RHdata and closed symbols with solid lines represent 2° C.-8° C. data.Data for lots A, B, C, D, E, and F are represented by circles (∘, ●),squares (□, ▪), triangles (Δ, ▴), diamonds (⋄,♦), stars (*) andhorizontal-lines (-), respectively. Values below limit of quantitation(LOQ) and not detected (ND) are shown as 0.05%, the LOQ.

FIG. 2B shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for 2-hydroxythiomorpholine(relative to cysteamine bitartrate; expressed as a percentage) insamples of 25-mg or 75-mg PROCYSBI®. Open symbols with dotted linesrepresent 25° C./60% RH data and closed symbols with solid linesrepresent 2° C.-8° C. data. Data for lots A, B, C, D, E and F arerepresented by circles (∘, ●), squares (□, ▪), triangles (Δ, ▴),diamonds (⋄,♦), stars (*) and horizontal-lines (-), respectively. Valuesbelow limit of quantitation (LOQ) and not detected (ND) are shown as0.05%, the LOQ.

FIG. 2C shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for cystamine tartrate amide(relative to cysteamine bitartrate; expressed as a percentage) insamples of 25-mg or 75-mg PROCYSBI®. Open symbols with dotted linesrepresent 25° C./60% RH data and closed symbols with solid linesrepresent 2° C.-8° C. data. Data for lots A, B, C, D, E and F arerepresented by circles (∘, ●), squares (□, ▪), triangles (Δ, ▴),diamonds (⋄,♦), stars (*) and horizontal-lines (-), respectively. Valuesbelow limit of quantitation (LOQ) and not detected (ND) are shown as0.05%, the LOQ.

FIG. 2D shows a comparison of cold storage (2° C.-8° C.) to roomtemperature storage (25° C./60% RH) for Peak 6 (relative to cysteaminebitartrate expressed as a percentage) in samples of 25-mg or 75-mgPROCYSBI®. Open symbols with dotted lines represent 25° C./60% RH dataand closed symbols with solid lines represent 2° C.-8° C. data. Data forlots A, B, C, D, E and F are represented by circles (∘, ●), squares (□,▪), triangles (Δ, ▴), diamonds (⋄,♦), stars (*) and horizontal-lines(-), respectively. Values below limit of quantitation (LOQ) and notdetected (ND) are shown as 0.05%, the LOQ.

FIG. 3A shows levels of cystamine (relative to cysteamine bitartrate;expressed as a percentage) for samples from Lot A (25-mg PROCYSBI®)after storage at 25° C./60% RH, 30° C./65% RH or 30° C./75% RH for 3months, following 15 months of initial storage at 2° C.-8° C. Closedcircles (●) with solid lines represent 2° C.-8° C. data. Open circles(∘) with dotted lines represent 25° C./60% RH data for 0-12 monthconsecutive storage as well as 15-18 months storage at 25° C./60% RHafter 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30°C./75% RH after 15 months storage at 2° C.-8° C.

FIG. 3B shows levels of cystamine (relative to cysteamine bitartrate;expressed as a percentage) for samples from Lot B (25-mg PROCYSBI®)after storage at 25° C./60% RH, 30° C./65% RH or 300/75% RH for 3months, following 15 months of initial storage at 2° C.-8° C. Closedcircles (●) with solid lines represent 2° C.-8° C. data. Open circles(∘) with dotted lines represent 25° C./60% RH data for 0-12 monthconsecutive storage as well as 15-18 months storage at 25° C./60% RHafter 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30°C./75% RH after 15 months storage at 2° C.-8° C.

FIG. 3C shows levels of 2-hydroxythiomorpholine (relative to cysteaminebitartrate; expressed as a percentage) for samples form Lot A (25-mgPROCYSBI®) after storage at 25° C./60% RH, 30° C./65% RH or 300/75% RHfor 3 months, following 15 months of initial storage at 2° C.-8° C.Closed circles (●) with solid lines represent 2° C.-8° C. data. Opencircles (∘) with dotted lines represent 25° C./60% RH data for 0-12month consecutive storage as well as 15-18 months storage at 25° C./60%RH after 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30°C./75% RH after 15 months storage at 2° C.-8° C.

FIG. 3D shows levels of 2-hydroxythiomorpholine (relative to cysteaminebitartrate; expressed as a percentage) for samples from Lot B (25-mgPROCYSBI®) after storage at 25° C./60% RH, 30° C./65% RH or 300/75% RHfor 3 months, following 15 months of initial storage at 2° C.-8° C.Closed circles (●) with solid lines represent 2° C.-8° C. data. Opencircles (∘) with dotted lines represent 25° C./60% RH data for 0-12month consecutive storage as well as 15-18 months storage at 25° C./60%RH after 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30°C./75% RH after 15 months storage at 2° C.-8° C.

FIG. 3E shows levels of cystamine tartrate amide (relative to cysteaminebitartrate; expressed as a percentage) for samples from Lot A (25-mgPROCYSBI®) after storage at 25° C./60% RH, 30° C./65% RH or 300/75% RHfor 3 months, following 15 months of initial storage at 2° C.-8° C.Closed circles (●) with solid lines represent 2° C.-8° C. data. Opencircles (∘) with dotted lines represent 25° C./60% RH data for 0-12month consecutive storage as well as 15-18 months storage at 25° C./60%RH after 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30°C./75% RH after 15 months storage at 2° C.-8° C.

FIG. 3F shows levels of cystamine tartrate amide (relative to cysteaminebitartrate; expressed as a percentage) for samples from Lot B (25-mgPROCYSBI®) after storage at 25° C./60% RH, 30° C./65% RH or 30°/75% RHfor 3 months, following 15 months of initial storage at 2° C.-8° C.Closed circles (●) with solid lines represent 2° C.-8° C. data. Opencircles (∘) with dotted lines represent 25° C./60% RH data for 0-12month consecutive storage as well as 15-18 months storage at 25° C./60%RH after 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30°C./75% RH after 15 months storage at 2° C.-8° C.

FIG. 3G shows levels of Peak 6 (relative to cysteamine bitartrate;expressed as a percentage) for samples from Lot A (25-mg PROCYSBI®)after storage at 25° C./60% RH, 30° C./65% RH or 300/75% RH for 3months, following 15 months of initial storage at 2° C.-8° C. Closedcircles (●) with solid lines represent 2° C.-8° C. data. Open circles(∘) with dotted lines represent 25° C./60% RH data for 0-12 monthconsecutive storage as well as 15-18 months storage at 25° C./60% RHafter 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30° °C./75% RH after 15 months storage at 2° C.-8° C.

FIG. 3H shows levels of Peak 6 (relative to cysteamine bitartrate;expressed as a percentage) for samples from Lot B (25-mg PROCYSBI®)after storage at 25° C./60% RH, 30° C./65% RH or 30° C./75% RH for 3months, following 15 months of initial storage at 2° C.-8° C. Closedcircles (●) with solid lines represent 2° C.-8° C. data. Open circles(∘) with dotted lines represent 25° C./60% RH data for 0-12 monthconsecutive storage as well as 15-18 months storage at 25° C./60% RHafter 15 months storage at 2° C.-8° C. Open squares (□) with dottedlines represent storage at 30° C./65% RH after 15 months storage at 2°C.-8° C. Open triangles (Δ) with dotted lines represent storage at 30°C./75% RH after 15 months storage at 2° C.-8° C.

DETAILED DESCRIPTION

In some aspects, the present disclosure provides methods of storingpharmaceutical compositions comprising cysteamine, or a pharmaceuticallyacceptable salt thereof, wherein the pharmaceutical composition isstored at refrigerated temperatures (e.g., 2° C.-8° C.) up to 24 months,or longer, and methods for using the same. In particular embodiments,pharmaceutical compositions handled according to the teachings providedherein provide greater chemical stability of cysteamine compared tocompositions stored at room temperature. More specifically, the relativeamounts of impurities in pharmaceutical compositions handled accordingto the present disclosure, measured at specific time points, are lessthan or equal to the corresponding amounts of impurities measured in thecompositions when stored at room temperature. Methods for distributingcysteamine compositions are also provided. The present disclosurefurther provides methods of treating cystinosis comprising administeringa pharmaceutical composition comprising cysteamine, or apharmaceutically acceptable salt thereof, wherein the pharmaceuticalcomposition has been stored under refrigerated temperatures prior toadministration.

Definitions

It should be understood that the terms “a” and “an” as used herein referto “one or more” of the enumerated components. The use of thealternative (e.g., “or”) should be understood to mean either one, both,or any combination thereof of the alternatives.

In addition, it should be understood that the individual features, orgroups of features, derived from the various combinations of themethods, pharmaceutical compositions, and substituents described herein,are disclosed by the present application to the same extent as if eachfeature or group of features was set forth individually. Thus, selectionof particular features is within the scope of the present disclosure.

As used herein, the terms “include.” “have,” and “comprise” are usedsynonymously, which terms and variants thereof are intended to beconstrued as non-limiting.

The term “consisting essentially of” limits the scope of a claim to thespecified materials or steps, or to those that do not materially affectthe basic characteristics of a claimed invention.

As used herein, the term “about” means±20% of the indicated range,value, or structure, unless otherwise indicated.

All ranges set forth herein include all possible subsets of ranges andany combinations of such subset ranges. Unless otherwise stated, rangesare inclusive of the stated endpoints. Where a range of values isprovided, it is understood that each intervening value between the upperand lower limit of that range and any other stated or intervening valuein that stated range, is encompassed within the disclosure. The upperand lower limits of these smaller ranges may independently be includedin the smaller ranges, and are also encompassed within the disclosure,subject to any specifically excluded limit in the stated range. Wherethe stated range includes one or both of the limits, ranges excludingeither or both of those included limits are also contemplated to be partof the disclosure. For example, “between 2 and 4” includes, but is notlimited to 2, 3, 4, 2 to 3, 3 to 4, and any number falling between 2 and4.

PROCYSBI® (cysteamine bitartrate; also known as ethanethiol, 2-amino,(2R,3R)-2,3-dihydroxybutanedioate) delayed-release capsules (RaptorPharmaceuticals, Inc.) is a prescription medicine used for the treatmentof nephropathic cystinosis. PROCYSBI® comprises enteric coatedcysteamine bitartrate beads encapsulated in gelatin capsules, andcurrently is available to patients in 25 mg and 75 mg strengths.PROCYSBI® contains the following inactive ingredients: microcrystallinecellulose, Eudragit® L 30 D-55, hypromellose, talc, triethyl citrate,sodium lauryl sulfate, and purified water.

An “enterically coated” drug or tablet refers to a drug, granule,granulation, powder or dosage form, including for example, a tablet, acaplet, and a capsule, that is coated with a substance—i.e., with an“enteric coating”—that remains intact in the stomach but dissolves andreleases the drug once the small intestine is reached.

As used herein “enteric coating”, is a material, a polymer material ormaterials which encase the medicament core (e.g., cysteamine,Cystagon®). Typically, a substantial amount or all of the entericcoating material is dissolved before the medicament or therapeuticallyactive agent is released from the dosage form, so as to achieve delayeddissolution of the medicament core. In one embodiment, a suitablepH-sensitive polymer is one which will dissolve in intestinal juices ata higher pH level (pH greater than 4.5), such as within the smallintestine and therefore permit release of the pharmacologically activesubstance in the regions of the small intestine, and not in the upperportion of the GI tract, such as the stomach.

By “pharmaceutically acceptable carrier” or “pharmaceutically acceptablevehicle” are meant materials that are suitable for oral administrationand not biologically, or otherwise, undesirable. i.e., that may beadministered to a subject along with an active ingredient withoutcausing any undesirable biological effects or interacting in adeleterious manner with any of the other components of a pharmaceuticalcomposition in which it is contained.

Similarly, a “pharmaceutically acceptable” salt, ester, or otherderivative of an active agent comprise, for example, salts, esters, orother derivatives which are not biologically or otherwise undesirable.

“Stabilizing agents” refer to compounds that lower the rate at which apharmaceutical product degrades, particularly an oral pharmaceuticalformulation under environmental conditions of storage.

By the terms “effective amount” or “therapeutically effective amount” ofa formulation of cysteamine refers to a nontoxic but sufficient amountof the agent to provide the desired therapeutic effect. The exact amountrequired will vary from subject to subject, depending on the age,weight, and general condition of the subject, the severity of thecondition being treated, and the like.

A “solid composition” as used herein refers to any solid-statecomposition that is, or can be made into, a solid pharmaceutical dosageform. Solid compositions include, for example, bulk powders, granulesand granulations (including coated granules), and dosage forms suitablefor oral administration to a subject, such as tablets or capsules,including compressed or extruded compositions. Moreover, the term,“solid” does not necessarily imply a complete absence of liquid orgaseous media. For example, solids can have various interstices, whichmay partially or fully fill with other gaseous and/or liquid media.Thus, the term “solid composition” includes compositions that aresuspended (i.e., remain at least partially, if not substantially,insoluble) in liquid media, such as syrups, elixirs, and the like.

“Storage” refers to maintaining a pharmaceutical composition under a setof physical conditions for a period of time. For example, storage caninclude maintaining a pharmaceutical composition at a particulartemperature, humidity, or both (e.g., 25° C./60% RH) for a givenduration (e.g., 4 weeks up to 24 months, or longer). As used herein,“storage” can include, for example, storage by a manufacturer, adistributor, a pharmacy, or a hospital prior to dispensing thepharmaceutical composition to a patient or health care provider.“Storage” can also include handling by a patient, wherein the patientmaintains a pharmaceutical composition under a set of physicalconditions for a period of time.

Addition definitions are set forth throughout this disclosure.

Methods of Storing and Stabilizing Pharmaceutical Compositions

In certain aspects, the present disclosure provides methods of storingor distributing pharmaceutical compositions comprising cysteamine, or apharmaceutically acceptable salt thereof, wherein the pharmaceuticalcompositions are stored under refrigerated conditions (e.g., 2° C.-8°C.). When compositions comprising cysteamine are stored at roomtemperature (e.g., 25° C.), impurities can increase over time ascysteamine degrades. The inventors have found that solid compositionsincluding cysteamine as an active ingredient (or agent) exhibitunexpectedly high levels of impurities associated with cysteaminedegradation when stored at 25° C./60% RH for 12 months. Impurities thatmay increase over time include cystamine, 2-hydroxythiomorpholine,cystamine tartrate amide, 2-hydroxymethylthiazolidine, and Peak 6. Incontrast, when cysteamine-containing compositions are stored attemperatures of about 2° C. to about 8° C. for 12 months, or even 24months, the level of each impurity is less than or equal to that foundin the compositions stored at 25° C./60% RH for 12 months.

In one embodiment, the methods as described herein relate to methodsinvolving pharmaceutical compositions comprising cysteamine, or apharmaceutically acceptable salt thereof, and one or more materials thatprovide increased delivery of cysteamine to the small intestine. In oneembodiment, a material that provides increased delivery of cysteamine tothe small intestine comprises an enteric coating. Typically, asubstantial amount of all of the enteric coating material is dissolvedbefore the medicament or therapeutically active agent is released fromthe dosage form, so as to achieve delayed dissolution of the medicamentcore. For example, a suitable pH-sensitive polymer is one which willdissolve in intestinal juices at a higher pH level (pH greater than4.5), such as within the small intestine, and therefore permit releaseof the pharmacologically active substance in the regions of the smallintestine and not in the upper portion of the GI tract, such as thestomach. In one embodiment, the enteric material begins to dissolve inan aqueous solution at pH between about 4.5 to about 5.5. In anotherembodiment, the enteric material begins to dissolve in an aqueoussolution at pH between about 5.5 to about 6.5. In another embodiment,the enteric material rapidly dissolves in an aqueous solution at pHbetween of about 5. In still another embodiment, the enteric materialrapidly dissolves in an aqueous solution at pH between of about 5.5. Inspecific embodiments, the cysteamine-containing composition may be adelayed release solid dosage form containing cysteamine, such as, forexample, PROCYSBI® (Raptor Pharmaceuticals, Inc.).

For example, pH-sensitive materials will not undergo significantdissolution until the dosage form has emptied from the stomach. The pHof the small intestine gradually increases from about 4.5 to about 6.5in the duodenal bulb to about 7.2 in the distal portions of the smallintestine (ileum). In order to provide predictable dissolutioncorresponding to the small intestine transit time of about 3 hours(e.g., 2-3 hours) and permit reproducible release therein, the membraneshould begin to dissolve within the pH range of the duodenum, andcontinue to dissolve at the pH range within the small intestine.Therefore, in one embodiment, the amount (thickness) of enteric membraneshould be sufficient to be substantially dissolved during theapproximate three hour transit time within the small intestine (e.g.,the proximal and mid-small intestine).

Enteric (gastro-resistant) materials can include, but are not limitedto, one or more of the following: cross-linked polyvinyl pyrrolidone;non-cross linked polyvinylpyrrolidone; hydroxypropylmethyl cellulosephthalate, hydroxypropylmethyl cellulose acetate succinate, celluloseacetate succinate; cellulose acetate phthalate, hydroxypropylmethylcellulose acetate succinate, cellulose acetate trimellitate; starchacetate phthalate; polyvinyl acetate phthalate; carboxymethyl cellulose;methyl cellulose phthalate; methyl cellulose succinate; methyl cellulosephthalate succinate; methyl cellulose phthalic acid half ester, ethylcellulose succinate; carboxymethylamide; potassiummethacrylatedvinylbenzene copolymer, polyvinylalcohols;polyoxyethyleneglycols; polyethylene glycol; sodium alginate;galactomannone; carboxypolymethylene; sodium carboxymethyl starch;copolymers of acrylic acid and/or methacrylic acid with a monomerselected from the following: methyl methacrylate, ethyl methacrylate,ethyl acrylate, butyl methacrylate, hexyl methacrylate, decylmethacrylate, lauryl methacrylate, phenyl methacrylate, methyl acrylate,isopropyl acrylate, isobutyl acrylate, or octadecyl acrylate, e.g.,EUDRAGIT-L and -S series, including L 100-55, L 30 D-55, L 100, S 100, L12.5, and S 12.5, available from Evonik Industries; polyvinyl acetate;fats; oils; waxes, fatty alcohols; shellac; zein; gluten;ethylacrylate-maleic acid anhydride copolymer; maleic acidanhydride-vinyl methyl ether copolymer; styrol-maleic acid copolymer;2-ethyl-hexyl-acrylate maleic acid anhydride; crotonic acid-vinylacetate copolymer; glutaminic acid/glutamic acid ester copolymer;carboxymethylethylcellulose glycerol monooctanoate; polyarginine;poly(ethylene); poly(propylene); poly(ethylene oxide); poly(ethyleneterephthalate); poly(vinyl isobutyl ether); poly(vinyl chloride); andpolyurethane. A combination of enteric materials may also be used. Inone embodiment, the enteric material rapidly dissolves at pH 5.5 andhigher, to provide fast dissolution in the upper bowel. For example, theenteric material can be selected from a copolymer of methacrylic acidand methyl methacrylate, and a copolymer of methacrylic acid and ethylacrylate. For example, an enteric polymer is poly(methacrylic acidco-ethyl acrylate) 1:1 (EUDRAGIT L 30 D-55 and EUDRAGIT L100-55).

Examples of some enteric coatings are disclosed in U.S. Pat. No.5,225,202, including beeswax and glyceryl monostearate; beeswax, shellacand cellulose; and cetyl alcohol, mastic and shellac, as well as shellacand stearic acid (U.S. Pat. No. 2,809,918); polyvinyl acetate and ethylcellulose (U.S. Pat. No. 3,835,221); and neutral copolymer ofpolymethacrylic acid esters (Eudragit L30D) (F. W. Goodhart et al.,Pharm. Tech., pp. 64-71, April 1984); copolymers of methacrylic acid andmethacrylic acid methylester (Eudragits), as a neutral copolymer ofpolymethacrylic acid esters containing metallic stearates (Mehta et al.,U.S. Pat. Nos. 4,728,512 and 4,794,001). Such coatings comprise mixturesof fats and fatty acids, shellac and shellac derivatives and thecellulose acid phthalates. e.g., those having a free carboxyl content.See also Remington's Pharmaceutical Sciences, A. Osol, ed., Mack Pub.Co., Easton, Pa. (16^(th) ed. 1980) at pages 1590-1593, and Zeitova etal. (U.S. Pat. No. 4,432,966), for descriptions of suitable entericcoating compositions.

One or more plasticizers can be added to enteric polymers in order toincrease their pliability and reduce brittleness, as it is known in theart. Suitable plasticizers are known in the art and include, forexample, butyl citrates, triethyl citrate, diethyl phthalate, dibutylsebacate, PEGs (e.g., PEG 6000), acetyl triethyl citrate, and triacetin.In one type of embodiment, the plasticizer is triethyl citrate. Whilesome enteric materials are flexible and do not require addition ofplasticizers, more brittle polymers (e.g., Eudragit US types. EudragitRIJRS, and Eudragit FS 30 D) benefit from plasticizers, e.g. in therange of 5 wt. % to 30 wt. % based on the dry polymer mass, e.g. about 8wt. % to about 12 wt. % triethyl citrate with poly(methacrylic acidco-ethyl acrylate) 1:1.

One or more anti-tacking agents (antiadherents) can also be added to anenteric coating mixture in order to reduce the tackiness of the film andprevent agglomeration, as it is known in the art. Anti-tacking agentsinclude talc, and glyceryl monostearate, fumed silica (e.g., AEROSIL200), precipitated silica (e.g., SIPERNAT PQ), and magnesium stearate,for example. Anti-tacking agents can be used in any suitable quantity,for example in a range of about 10 wt. % to 100 wt. % based on drypolymer mass, or about 10 wt. % to about 50 wt. %, or about 10 wt. % toabout 30 wt. %, or about 15 wt. % to about 30 wt. %. For example, in oneembodiment the amount of talc is in a range of 15 wt. % to about 30 wt.%, based on dry polymer mass.

One or more surfactants can also be added to an enteric coating mixturein order to improve substrate wettability and/or stabilize suspensions,as it is known in the art. Surfactants include Polysorbate 80, sorbitanmonooleate, and sodium dodecyl sulfate, for example.

The enteric membrane can be formed by any suitable process. Coatingprocesses include pan coating, fluid bed coating, and dry coating (e.g.,heat dry coating and electrostatic dry coating), for example. Pancoating and fluid bed coating using solvent are well establishedprocesses. In liquid coating, the enteric material and optionalexcipients (e.g., pigments, plasticizers, anti-tacking agents) are mixedin an organic solvent or water to form a solution or dispersion. Thecoating solution or dispersion is sprayed into solid dosage forms in apan coater or a fluid bed dryer and dried by hot air. For example, in aWurster fluid bed coating process, the coating fluid is sprayed from thebottom of the fluid bed apparatus, whereas in an alternative the coatingfluid is applied by top spraying, and in another alternative tangentialspray is applied.

The amount of enteric material applied is sufficient to achieve desiredacid resistance and release characteristics. For example, in oneembodiment the amount of enteric membrane will be sufficient to meetUnited States Pharmacopeia (USP)<711> requirements (USP 36-NF 31) fordelayed-release dosage forms, thereby not releasing 10.0 wt. % drugafter 2 hours in 0.1N HCl. In another aspect, a formulation will besufficient to release at least 80% of the active in 20 minutes in pH 6.8buffer solution, e.g., using the dissolution method of USP 36-NF 31section <711>.

In one embodiment, an enteric membrane is present in an amount in arange of about 20% to 40%, or 25% to about 35% as measured by the weightgain compared to the uncoated particle cores, or in a range of about 25%to about 31% weight gain, or about 27% to about 31% weight gain, orabout 28.5% to about 31% weight gain, based on the weight of theuncoated particle cores.

In pharmaceutical compositions used in the methods described herein, thecysteamine is present in the compositions in a therapeutically effectiveamount; in one embodiment, the composition is in unit dosage form. Theamount of cysteamine administered will be dependent on the age, weight,and general condition of the subject, the severity of the conditionbeing treated, and the judgment of the prescribing physician. In oneembodiment, the dose is administered twice per day at about 0.5-1.0 g/m²(e.g., 0.7-0.8 g/m²) body surface area. Current non-enterically coateddoses are about 1.35 g/m² body surface area and are administered 4-5times per day. In another embodiment, the dose is about 0.2-1.95 g/m²body surface area.

In one aspect, a method of storing a pharmaceutical composition isprovided, comprising storing the pharmaceutical composition at atemperature of between about 2° C. and about 8° C., wherein thepharmaceutical composition comprises cysteamine, or a pharmaceuticallyacceptable salt thereof. In another aspect, a method of stabilizing apharmaceutical composition is provided, comprising storing thepharmaceutical composition at a temperature of between about 2° C. andabout 8° C. wherein the pharmaceutical composition comprises cysteamine,or a pharmaceutically acceptable salt thereof.

In any of the aforementioned methods, the pharmaceutical composition mayfurther comprise one or more materials that provide increased deliveryof cysteamine to the small intestine. For example, in one embodiment,the material that provides increased delivery of cysteamine to the smallintestine comprises an enteric coating, such as a coating selected fromthe group consisting of polymerized gelatin, shellac, methacrylic acidcopolymer type CNF, cellulose butyrate phthalate, cellulose hydrogenphthalate, cellulose proprionate phthalate, polyvinyl acetate phthalate(PVAP), cellulose acetate phthalate (CAP), cellulose acetatetrimellitate (CAT), hydroxypropyl methylcellulose phthalate,hydroxypropyl methylcellulose acetate, dioxypropyl methylcellulosesuccinate, carboxymethyl ethylcellulose (CMEC), hydroxypropylmethylcellulose acetate succinate (HPMCAS), and acrylic acid polymersand copolymers formed from methyl acrylate, ethyl acrylate, methylmethacrylate, and/or ethyl methacrylate with copolymers of acrylic andmethacrylic acid esters. In one embodiment, the enteric coatingcomprises poly(methacrylic acid co-ethyl acrylate) 1:1 (Eudragit L30-D-55).

In any of the aforementioned embodiments, the pharmaceutical compositionmay comprise a pharmaceutically acceptable salt of cysteamine. In oneembodiment, the pharmaceutically acceptable salt of cysteamine iscysteamine bitartrate.

In any of the aforementioned embodiments, the pharmaceutical compositionmay comprise a solid composition. In one embodiment, the pharmaceuticalcomposition comprises a unit dose of about 25 mg cysteamine. In anotherembodiment, the pharmaceutical composition comprises a unit dose ofabout 75 cysteamine. Exemplary pharmaceutical compositions comprisingcysteamine are disclosed in International Patent Publication WO2014/204881.

In various embodiments, the pharmaceutical composition comprises entericcoated beads. In various embodiments, the pharmaceutical composition isa pharmaceutical dosage form that includes a plurality of cysteaminebeads, the beads including a core particle including cysteamine or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable excipient, and an enteric membrane surrounding the coreparticle, wherein the plurality of beads is characterized by adistribution of particle sizes.

In one embodiment, the particle sizes of the beads are in a range ofabout 0.7 mm to about 2.5 mm, or about 0.7 mm to about 2.8 mm, or about0.8 mm to about 1.7 mm. For example, the target bead size can be up to2.5 mm with no more than 10 percent variation over this size, to amaximum size of 2.8 mm. In one embodiment, the enteric membrane on thebeads is present in an amount in a range of about 20% to 40%, or about20% to about 25%, or about 25% to about 35% as measured by the weightgain compared to the uncoated particle cores, or in a range of about 25%to about 31% weight gain, or about 27% to about 31% weight gain, orabout 28.5% to about 31% weight gain, based on the weight of theuncoated particle cores.

In various embodiments, the bead formulation comprises a plurality ofcysteamine beads, the beads comprising a core particle comprisingcysteamine, optionally cysteamine bitartrate, a filler, a binder and anenteric membrane surrounding the core, wherein the plurality of beads ischaracterized by a distribution of particle sizes in a range of about0.7 mm to about 2.8 mm; wherein the enteric membrane is present in anamount in a range of about 20% to about 40% based on the weight of thebead core particles; wherein the formulation is a delayed releaseformulation having an enteric membrane that begins to dissolve within apH range of about 4.5 to about 6.5, and wherein the beads are disposedin a capsule shell.

In one embodiment, a method of storing a pharmaceutical composition isprovided, comprising storing the pharmaceutical composition at atemperature of between about 2° C. and about 8° C., wherein thepharmaceutical composition comprises cysteamine bitartrate. In a furtherembodiment, the pharmaceutical composition comprises PROCYSBI®. Inanother embodiment, a method of stabilizing a pharmaceutical compositionis provided, comprising storing the pharmaceutical composition at atemperature of between about 2° C. and about 8° C. wherein thepharmaceutical composition comprises cysteamine bitartrate. In a furtherembodiment, the pharmaceutical composition comprises PROCYSBI®.

In any of the aforementioned embodiments, the pharmaceutical compositionmay be stored at a temperature of between about 2° C. and about 8° C.for up to 1 month, between about 1 month and about 6 months, betweenabout 6 months and about 12 months, between about 12 months and about 15months, between about 15 months and about 18 months, between about 18months and about 21 months, between about 21 months and about 24 months,between about 24 months and about 36 months, or between about 36 monthsand about 39 months. In one embodiment, the pharmaceutical compositionis stored at a temperature of between about 2° C. and about 8° C. for 1month, 6 months, 12 months, 15 months, 18 months, 21 months, 24 months,36 months, or 39 months.

In any of the aforementioned embodiments, the method may furthercomprise storing the pharmaceutical composition at a temperature ofbetween about 20° C. and about 25° C. and a relative humidity of 60% forup to 4 months.

In any of the aforementioned embodiments, the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan 0.5%. For example, in one embodiment, a method of storing apharmaceutical composition is provided, wherein (a) the pharmaceuticalcomposition is stored at a temperature of between about 2° C. and about8° C. for up to 24 months. (b) the pharmaceutical composition comprisescysteamine bitartrate, and (c) the amount of 2-hydroxythiomorpholinepresent in the pharmaceutical composition, relative to the amount ofcysteamine bitartrate present in the pharmaceutical composition, is lessthan 0.5%. In another embodiment, a method of storing or stabilizing apharmaceutical composition is provided, wherein the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, is less than orequal to the amount of 2-hydroxythiomorpholine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25° C. and a relative humidity of 60% for the same duration.

In any one of the aforementioned embodiments, the amount of cystaminepresent in the pharmaceutical composition, relative to the amount ofcysteamine, or a pharmaceutically acceptable salt thereof, present inthe pharmaceutical composition, may be less than 4%. For example, in oneembodiment, a method of storing a pharmaceutical composition isprovided, wherein (a) the pharmaceutical composition is stored at atemperature of between about 2° C. and about 8° C. for up to 24 months,(b) the pharmaceutical composition comprises cysteamine bitartrate, and(c) the amount of cystamine present in the pharmaceutical composition,relative to the amount of cysteamine bitartrate present in thepharmaceutical composition, is less than 4%. In another embodiment, amethod of storing or stabilizing a pharmaceutical composition isprovided, wherein the amount of cystamine present in the pharmaceuticalcomposition, relative to the amount of cysteamine, or a pharmaceuticallyacceptable salt thereof, present in the pharmaceutical composition, isless than or equal to the amount of cystamine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25° C. and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the amount of cystaminetartrate amide present in the pharmaceutical composition, relative tothe amount of cysteamine, or a pharmaceutically acceptable salt thereof,present in the pharmaceutical composition, may be less than 0.5%. Forexample, in one embodiment, a method of storing a pharmaceuticalcomposition is provided, wherein (a) the pharmaceutical composition isstored at a temperature of between about 2° C. and about 8° C. for up to24 months, (b) the pharmaceutical composition comprises cysteaminebitartrate, and (c) the amount of cystamine tartrate amide present inthe pharmaceutical composition, relative to the amount of cysteaminebitartrate present in the pharmaceutical composition, is less than 0.5%.In another embodiment, a method of storing or stabilizing apharmaceutical composition is provided, wherein the amount of cystaminetartrate amide present in the pharmaceutical composition, relative tothe amount of cysteamine, or a pharmaceutically acceptable salt thereof,present in the pharmaceutical composition, is less than or equal to theamount of cystamine tartrate amide present in the pharmaceuticalcomposition, relative to the amount of cysteamine, or a pharmaceuticallyacceptable salt thereof, present in the pharmaceutical composition,after storage at a temperature of between about 20° C. and about 25° C.and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the amount of2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan 0.5%. For example, in one embodiment, a method of storing apharmaceutical composition is provided, wherein (a) the pharmaceuticalcomposition is stored at a temperature of between about 2° C. and about8° C. for up to 24 months, (b) the pharmaceutical composition comprisescysteamine bitartrate, and (c) the amount of 2-hydroxymethylthiazolidinepresent in the pharmaceutical composition, relative to the amount ofcysteamine bitartrate present in the pharmaceutical composition, is lessthan 0.5%. In another embodiment, a method of storing or stabilizing apharmaceutical composition is provided, wherein the amount of2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, is less than orequal to the amount of 2-hydroxymethylthiazolidine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25° C. and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the total amount of2-hydroxythiomorpholine, cystamine, cystamine tartrate amide, and2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan or equal to the total amount of 2-hydroxythiomorpholine, cystamine,cystamine tartrate amide, and 2-hydroxymcthylthiazolidine present in thepharmaceutical composition, relative to the amount cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25° C. and a relative humidity of 60% for the same duration.

Methods of Distributing Pharmaceutical Compositions

In another aspect, a method of distributing a pharmaceutical compositionis provided. In one embodiment, the method comprises storing thepharmaceutical composition at a temperature of between about 2° C. andabout 8° C. prior to dispensing to a health care provider or a patient,wherein the pharmaceutical composition comprises cysteamine, or apharmaceutically acceptable salt thereof. In another embodiment, apharmaceutical composition comprising cysteamine, or a pharmaceuticallyacceptable salt thereof, is shipped from a manufacture to a distributorunder refrigerated conditions, and is then stored by the distributorprior to dispensing to a health care provider or a patient. In oneembodiment, a pharmaceutical composition comprising cysteamine, or apharmaceutically acceptable salt thereof, is stored by a distributor ata temperature of between about 2° C. and about 8° C. for between about 4weeks and about 24 months, or longer.

In any of the aforementioned methods of distributing, the pharmaceuticalcomposition may further comprise one or more materials that provideincreased delivery of cysteamine to the small intestine. For example, inone embodiment, the material that provides increased delivery ofcysteamine to the small intestine comprises an enteric coating, such asa coating selected from the group consisting of polymerized gelatin,shellac, methacrylic acid copolymer type CNF, cellulose butyratephthalate, cellulose hydrogen phthalate, cellulose proprionatephthalate, polyvinyl acetate phthalate (PVAP), cellulose acetatephthalate (CAP), cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate, hydroxypropyl methylcellulose acetate,dioxypropyl methylcellulose succinate, carboxymethyl ethylcellulose(CMEC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), andacrylic acid polymers and copolymers formed from methyl acrylate, ethylacrylate, methyl methacrylate, and/or ethyl methacrylate with copolymersof acrylic and methacrylic acid esters. In one embodiment, the entericcoating comprises poly(methacrylic acid co-ethyl acrylate) 1:1 (EudragitL 30-D-55).

In any of the aforementioned embodiments, the pharmaceutical compositionmay comprise a pharmaceutically acceptable salt of cysteamine. In oneembodiment, the pharmaceutically acceptable salt of cysteamine iscysteamine bitartrate.

In any of the aforementioned embodiments, the pharmaceutical compositionmay comprise a solid composition. In one embodiment, the pharmaceuticalcomposition comprises a unit dose of about 25 mg cysteamine. In anotherembodiment, the pharmaceutical composition comprises a unit dose ofabout 75 mg cysteamine.

In one embodiment, a method of distributing a pharmaceutical compositionis provided, comprising storing the pharmaceutical composition at atemperature of between about 2° C. and about 8° C. prior to dispensingto a health care provider or a patient, wherein the pharmaceuticalcomposition comprises cysteamine bitartrate. In a further embodiment,the pharmaceutical composition comprises PROCYSBI®.

In any of the aforementioned embodiments, the pharmaceutical compositionmay be stored at a temperature of between about 2° C. and about 8° C.for up to 1 month, between about 1 month and about 6 months, betweenabout 6 months and about 12 months, between about 12 months and about 15months, between about 15 months and about 18 months, between about 18months and about 21 months, between about 21 months and about 24 months,between about 24 months and about 36 months, or between about 36 monthsand about 39 months. In one embodiment, the pharmaceutical compositionis stored at a temperature of between about 2° C. and about 8° C. for 1month, 6 months, 12 months, 15 months, 18 months, 21 months, 24 months,36 months, or 39 months.

In any of the aforementioned embodiments, the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan 0.5%. In another embodiment, a method of distributing apharmaceutical composition is provided, wherein the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, is less than orequal to the amount of 2-hydroxythiomorpholine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25° C. and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the amount of cystaminepresent in the pharmaceutical composition, relative to the amount ofcysteamine, or a pharmaceutically acceptable salt thereof, present inthe pharmaceutical composition, may be less than 4%. In anotherembodiment, a method of distributing a pharmaceutical composition isprovided, wherein the amount of cystamine present in the pharmaceuticalcomposition, relative to the amount of cysteamine, or a pharmaceuticallyacceptable salt thereof, present in the pharmaceutical composition, isless than or equal to the amount of cystamine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25 C.° and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the amount of cystaminetartrate amide present in the pharmaceutical composition, relative tothe amount of cysteamine, or a pharmaceutically acceptable salt thereof,present in the pharmaceutical composition, may be less than 0.5%. Inanother embodiment, a method of distributing a pharmaceuticalcomposition is provided, wherein the amount of cystamine tartrate amidepresent in the pharmaceutical composition, relative to the amount ofcysteamine, or a pharmaceutically acceptable salt thereof, present inthe pharmaceutical composition, is less than or equal to the amount ofcystamine tartrate amide present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, after storageat a temperature of between about 20° C. and about 25° C. and a relativehumidity of 60% for the same duration.

In any of the aforementioned embodiments, the amount of2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan 0.5%. In another embodiment, a method of distributing apharmaceutical composition is provided, wherein the amount of2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, is less than orequal to the amount of 2-hydroxymethylthiazolidine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25° C. and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the total amount of2-hydroxythiomorpholine, cystamine, cystamine tartrate amide, and2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan or equal to the total amount of 2-hydroxythiomorpholine, cystamine,cystamine tartrate amide, and 2-hydroxymethylthiazolidine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 200 andabout 25° and a relative humidity of 60% for the same duration.

Method of Treatment

In another aspect, the present disclosure provides methods of treating adisease or disorder by administering to a subject in need thereof apharmaceutical composition comprising cysteamine, or a pharmaceuticallyacceptable salt thereof, wherein the pharmaceutical composition has beenstored at a temperature of between about 2° C. and about 8° C. prior toadministration. In various embodiments, the disease or disorder iscystinosis, fatty liver disease, a thrombotic disease, an MECP-2 relateddisorder, an inherited mitochondrial disease, a neurological disease ordisorder, inflammation and cancer.

In various embodiments, the fatty liver disease is selected from thegroup consisting of non-alcoholic fatty liver disease (NAFLD),non-alcoholic steatohepatitis (NASH), fatty liver disease resulting fromhepatitis, fatty liver disease resulting from obesity, fatty liverdisease resulting from diabetes, fatty liver disease resulting frominsulin resistance, fatty liver disease resulting fromhypertriglyceridemia, Abetalipoproteinemia, glycogen storage diseases.Weber-Christian disease, Wolmans disease, acute fatty liver ofpregnancy, and lipodystrophy.

In various embodiments, the thrombotic disease is selected from thegroup consisting of sickle cell disease, deep vein thrombosis, pulmonaryembolism, cardiac embolism, hypercoagulable state, thrombophilia, FactorV Leiden, Antithrombin III deficiency, Protein C deficiency. Protein Sdeficiency. Prothrombin gene mutation (G20210A), Hyperhomcysteinemia,antiphospholipid antibody syndrome (APS), anticardiolipin antibody(ACLA) thrombosis syndrome, or lupus anticoagulant (LA) syndrome.

In various embodiments, the neurological disease or disorder is selectedfrom the group consisting of Huntington's Disease, Parkinson's Disease,amyotrophic lateral sclerosis, multiple sclerosis. Alzheimer's diseasespinal muscle atrophy, concussion, stroke, and traumatic brain injury(TBI).

In various embodiments, the MECP-2 related disease is selected from thegroup consisting of Rett syndrome, autism, pervasive developmentdisorder, non-syndromic mental retardation, idiopathic neonatalencephalopathy and idiopathic cerebral palsy.

In various embodiments, the inherited mitochondrial disease is selectedfrom the group consisting of Friedreich's ataxia, Leber's hereditaryoptic neuropathy (LHON), myoclonic epilepsy and ragged-red fibers,mitochondrial encephalomyopathy, lactic acidosis, and stroke-likesyndrome (MELAS). Keam-Sayre syndrome and subacute necrotizingencephalopathy (Leigh's Syndrome).

In various embodiments, the cancer is selected from the group consistingof breast cancer, melanoma, prostate cancer, pancreatic cancer, head andneck cancer, lung cancer, non small-cell lung carcinoma, renal cancer,colorectal cancer, colon cancer, ovarian cancer, liver cancer andgastric cancer.

In one aspect, the cysteamine composition is administered in dosageform, wherein the dose is administered either one time per day ormultiple times per day. The cysteamine composition may be administeredless than four times per day, e.g., one, two or three times per day. Insome embodiments, an effective dosage of cysteamine composition may bewithin the range of 0.01 mg to 1000 mg per kg (mg/kg) of body weight perday. Further, the effective dose may be 0.5 mg/kg, 1 mg/kg, 5 mg/kg, 10mg/kg, 15 mg/kg, 20 mg/kg/25 mg/kg, 30 mg/kg, 35 mg/kg, 40 mg/kg, 45mg/kg, 50 mg/kg, 55 mg/kg, 60 mg/kg, 70 mg/kg, 75 mg/kg, 80 mg/kg, 90mg/kg, 100 mg/kg, 125 mg/kg, 150 mg/kg, 175 mg/kg, 200 mg/kg, 225 mg/kg,250 mg/kg, 275 mg/kg, 300 mg/kg, 325 mg/kg, 350 mg/kg, 375 mg/kg, 400mg/kg, 425 mg/kg, 450 mg/kg, 475 mg/kg, 500 mg/kg, 525 mg/kg, 550 mg/kg,575 mg/kg, 600 mg/kg, 625 mg/kg, 650 mg/kg, 675 mg/kg, 700 mg/kg, 725mg/kg, 750 mg/kg, 775 mg/kg, 800 mg/kg, 825 mg/kg, 850 mg/kg, 875 mg/kg,900 mg/kg, 925 mg/kg, 950 mg/kg, 975 mg/kg or 1000 mg/kg, or may rangebetween any two of the foregoing values. In some embodiments, the doseabove may be the total daily dose, or may be the dose administered inone of the one, two or three daily administrations. In some embodiments,the cysteamine composition is administered at a total daily dose of fromapproximately 0.25 g/m2 to 4.0 g/m2 body surface area, e.g., at leastabout 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7,1.8, 1.9 or 2 g/m2, or up to about 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4,1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.2, 2.5, 2.7, 3.0, or 3.5 g/m2 or mayrange between any two of the foregoing values. In some embodiments, thecysteamine composition may be administered at a total daily dose ofabout 0.5-2.0 g/m² body surface area, or 1-1.5 g/m² body surface area,or 0.5-1 g/m² body surface area, or about 0.7-0.8 g/m² body surfacearea, or about 1.3 g/m² body surface area (e.g., about 1.35 g/m² bodysurface area), or about 1.3 to about 1.95 grams/m²/day, or about 0.5 toabout 1.5 grams/m²/day, or about 0.5 to about 1.0 grams/m²/day,preferably at a frequency of fewer than four times per day, e.g. three,two or one times per day. Salts or esters of the same active ingredientmay vary in molecular weight depending on the type and weight of thesalt or ester moiety. For administration of enteric dosage form, e.g., atablet or capsule or other oral dosage form comprising the entericallycoated cysteamine product, a total weight in the range of approximately100 mg to 1000 mg is used. In certain embodiments, the amount ofcysteamine active ingredient in a tablet or capsule is approximately 15,20, 25, 50, 75, 100, 125, 150, 175, 200, 250, 300, 400 or 500 mg.

In one embodiment, provided is a method of treating cystinosis,comprising administering a therapeutically effective amount of apharmaceutical composition comprising cysteamine, or a pharmaceuticallyacceptable salt thereof, wherein the pharmaceutical composition has beenstored at a temperature of between about 2° C. and about 8° C. prior toadministration.

In various embodiments, the pharmaceutical composition is formulated toprovide white blood cell cystine suppression with a 12 hour level below1 nmol cystine/mg protein.

In various embodiments, each dose of cysteamine is about 0.5-1.0 g/m2body surface area. In various embodiments, the total daily dose ofcysteamine is about 1.30 g/m2 body surface area or less.

In various embodiments, the composition increases delivery to theproximal small intestine, the mid-small intestine, the duodenum, thejejunum or the mid-ileum.

In various embodiments, the composition is in the form of a tablet or acapsule.

In various embodiments, the cysteamine salt is cysteamine bitartrate.

The methods of treatment of the disclosure can be used in combinationwith other therapies useful for treating cystinosis andneurodegenerative diseases and disorders. For example, indomethacintherapy (Indocid® or Endol®) is an anti-inflammatory used to treatrheumatoid arthritis and lumbago, but it can be used to reduce water andelectrolyte urine loss. In children with cystinosis, indomethacinreduces the urine volume and therefore liquid consumption by about 30%,sometimes by half. In most cases this is associated with an appetiteimprovement. Indomethacin treatment is generally followed for severalyears.

Other therapies can be combined with the methods and compositions of thedisclosure to treat diseases and disorders that are attributed to orresult from cystinosis. Urinary phosphorus loss, for example, entailsrickets, and it may be necessary to give a phosphorus supplement.Carnitine is lost in the urine and blood levels are low. Carnitineallows fat to be used by the muscles to provide energy. Hormonesupplementation is sometimes necessary. Sometimes the thyroid gland willnot produce enough thyroid hormones. This is given as thyroxin (drops ortables). Insulin treatment is sometimes necessary if diabetes appears,when the pancreas does not produce enough insulin. These treatments havebecome rarely necessary in children whom are treated with cysteamine,since the treatment protects the thyroid and the pancreas. Someadolescent boys require a testosterone treatment if puberty is late.Growth hormone therapy may be indicated if growth is not sufficientdespite a good hydro electrolytes balance. Accordingly, such therapiescan be combined with the enterically coated cysteamine and cystaminecompositions and methods of the disclosure.

The effectiveness of a method of the disclosure can be assessed bymeasuring leukocyte cystine concentrations. Dosage adjustment andtherapy can be made by a medical specialist depending upon, for example,the severity of cystinosis and/or the concentration of cystine.Additional therapies including the use of omeprazole (Prilosec®) canreduce these symptoms.

Accordingly, in one embodiment, the present disclosure provides a methodof treating cystinosis, comprising administering a pharmaceuticalcomposition comprising cysteamine, or a pharmaceutically acceptable saltthereof, wherein the pharmaceutical composition has been stored at atemperature of between about 2° C. and about 8° C. prior toadministration. In another embodiment, a method of treating cystinosisis provided, wherein the pharmaceutical composition has been firststored at a temperature of between about 2° C. and about 8° C. prior toadministration and is subsequently stored at a temperature of betweenabout 20° C. and about 25° C. and a relative humidity of 60% for up to 4months prior to administration.

In any of the aforementioned methods of treatment, the pharmaceuticalcomposition may further comprise one or more materials that provideincreased delivery of cysteamine to the small intestine. For example, inone embodiment, the material that provides increased delivery ofcysteamine to the small intestine comprises an enteric coating, such asa coating selected from the group consisting of polymerized gelatin,shellac, methacrylic acid copolymer type CNF, cellulose butyratephthalate, cellulose hydrogen phthalate, cellulose proprionatephthalate, polyvinyl acetate phthalate (PVAP), cellulose acetatephthalate (CAP), cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate, hydroxypropyl methylcellulose acetate,dioxypropyl methylcellulose succinate, carboxymethyl ethylcellulose(CMEC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), andacrylic acid polymers and copolymers formed from methyl acrylate, ethylacrylate, methyl methacrylate, and/or ethyl methacrylate with copolymersof acrylic and methacrylic acid esters. In one embodiment, the entericcoating comprises poly(methacrylic acid co-ethyl acrylate) 1:1 (EudragitL 30-D-55).

In any of the aforementioned embodiments, the pharmaceutical compositionmay comprise a pharmaceutically acceptable salt of cysteamine. In oneembodiment, the pharmaceutically acceptable salt of cysteamine iscysteamine bitartrate.

In any of the aforementioned embodiments, the pharmaceutical compositionmay comprise a solid composition. In one embodiment, the pharmaceuticalcomposition comprises a unit dose of about 25 mg cysteamine. In anotherembodiment, the pharmaceutical composition comprises a unit dose ofabout 75 mg cysteamine. In various embodiments, the pharmaceuticalcomposition is a bead formulation as described herein.

In one embodiment, a method of treating cystinosis is provided,comprising administering a pharmaceutical composition comprisingcysteamine bitartrate, wherein the pharmaceutical composition has beenstored at a temperature of between about 2° C. and about 8° C. prior toadministration. In another embodiment, a method of treating cystinosisis provided, comprising administering a pharmaceutical compositioncomprising cysteamine bitartrate, wherein the pharmaceutical compositionhas been first stored at a temperature of between about 2° C. and about8° C. prior to administration and subsequently stored at a temperatureof between about 20° C. and about 25° C. and a relative humidity of 60%for up to 4 months prior to administration. In further embodiment, thepharmaceutical composition comprises PROCYSBI®.

In any of the aforementioned embodiments, the pharmaceutical compositionmay be stored at a temperature of between 2° C. and about 8° C. for upto 1 month, between about 1 month and about 6 months, between about 6months and about 12 months, between about 12 months and about 15 months,between about 15 months and about 18 months, between about 18 months andabout 21 months, between about 21 months and about 24 months, betweenabout 24 months and about 36 months, or between about 36 months andabout 39 months. In one embodiment, the pharmaceutical composition isstored at a temperature of between about 2° C. and about 8° C. for 1month, 6 months, 12 months, 15 months, 18 months, 21 months, 24 months,36 months, or 39 months.

In any of the aforementioned embodiments, the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan 0.5%. In another embodiment, a method of treating a cystinosis isprovided, wherein the amount of 2-hydroxythiomorpholine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, is less than or equal to the amount of2-hydroxythiomorpholine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, after storageat a temperature of between about 20° C. and about 25° C. and a relativehumidity of 60% for the same duration.

In any of the aforementioned embodiments, the amount of cystaminepresent in the pharmaceutical composition, relative to the amount ofcysteamine, or a pharmaceutically acceptable salt thereof, present inthe pharmaceutical composition, may be less than 4%. In anotherembodiment, a method of treating cystinosis is provided, wherein theamount of cystamine present in the pharmaceutical composition, relativeto the amount of cysteamine, or a pharmaceutically acceptable saltthereof, present in the pharmaceutical composition, is less than orequal to the amount of cystamine present in the pharmaceuticalcomposition, relative to the amount of cysteamine, or a pharmaceuticallyacceptable salt thereof, present in the pharmaceutical composition,after storage at a temperature of between about 20° C. and about 25° C.and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the amount of cystaminetartrate amide present in the pharmaceutical composition, relative tothe amount of cysteamine, or a pharmaceutically acceptable salt thereof,present in the pharmaceutical composition, may be less than 0.5%. Inanother embodiment, a method of treating cystinosis is provided, whereinthe amount of cystamine tartrate amide present in the pharmaceuticalcomposition, relative to the amount of cysteamine, or a pharmaceuticallyacceptable salt thereof, present in the pharmaceutical composition, isless than or equal to the amount of cystamine tartrate amide present inthe pharmaceutical composition, relative to the amount of cysteamine, ora pharmaceutically acceptable salt thereof, present in thepharmaceutical composition, after storage at a temperature of betweenabout 20° C. and about 25° C. and a relative humidity of 60% for thesame duration.

In any of the aforementioned embodiments, the amount of2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan 0.5%. In another embodiment, a method of treating cystinosis isprovided, wherein the amount of 2-hydroxymethylthiazolidine present inthe pharmaceutical composition, relative to the amount of cysteamine, ora pharmaceutically acceptable salt thereof, present in thepharmaceutical composition, is less than or equal to the amount of2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, after storageat a temperature of between about 20° C. and about 25° C. and a relativehumidity of 60% for the same duration.

In any of the aforementioned embodiments, the total amount of2-hydroxythiomorpholine cystamine, cystamine tartrate amide, and2-hydroxymethylthiazolidine present in the pharmaceutical composition,relative to the amount of cysteamine, or a pharmaceutically acceptablesalt thereof, present in the pharmaceutical composition, may be lessthan or equal to the total amount of 2-hydroxythiomorpholine, cystamine,cystamine tartrate amide, and 2-hydroxymethylthiazolidine present in thepharmaceutical composition, relative to the amount of cysteamine, or apharmaceutically acceptable salt thereof, present in the pharmaceuticalcomposition, after storage at a temperature of between about 20° C. andabout 25° C. and a relative humidity of 60% for the same duration.

In any of the aforementioned embodiments, the cystinosis may benephropathic cystinosis.

In another aspect, the present disclosure provides methods of treating adisease or disorder, comprising administering a therapeuticallyeffective amount of a pharmaceutical composition comprising cysteamine,or a pharmaceutical acceptable salt thereof, wherein the pharmaceuticalcomposition has been stored at a temperature of between about 2° C. andabout 8° C. prior to administration. In another embodiment, a method oftreating a disease or disorder is provided, wherein the pharmaceuticalcomposition has been first stored at a temperature of between about 2°C. and about 8° C. prior to administration and is subsequently stored ata temperature of between about 20° C. and about 25° C. and a relativehumidity of 60% for up to 4 months prior to administration. In variousembodiments, the disease or disorder is cystinosis, fatty liver disease,a thrombotic disease, an MECP-2 related disorder, an inheritedmitochondrial disease, a neurological disease or disorder, inflammationand cancer. In one embodiment, the disease or disorder is a metabolicand neurodegenerative disease, such as non-alcoholic fatty liver disease(NAFLD), non-alcoholic steatohepatitis (NASH), Huntington's disease,Parkinson's disease, Rett Syndrome. or cystic fibrosis. In anotherembodiment, the disease or disorder is a fibrotic disorder or aninflammatory disorder.

EXAMPLES Example 1 Impurity Levels in PROCYSBI® after Storage at 25° C.

Nineteen impurities have been identified for PROCYSBI® (Table 1). Sevenof these impurities do not increase during storage at 25° C./60%relative humidity (RH): Peak 3, Peak E. RRT 1.41-1.47, Peak F, Peak G,Peak H, and Peak I. Twelve impurities may increase when stored at 25°C./60% RH: cystamine, Peak A, Peak B, Peak C, 2-hydroxythiomorpholine,Peak D, cystamine tartrate amide, 2-hydroxymethylthiazolidine, Peak 5,Peak 6, Peak J, and Peak K.

TABLE 1 Impurities That Do and Do Not Increase on Stability When Storedat 25° C./60% RH Impurities that do not increase Impurities that mayincrease when stored at 25° C./60% RH when stored at 25° C./60% RH Peak3 Cystamine Peak E Peak A RRT 1.41-1.47 Peak B Peak F Peak C Peak G2-hydroxythiomorpholine Peak H Peak D Peak I Cystamine tartrate amide2-hydroxymethylthiazolidine Peak 5 Peak 6 Peak J Peak K

Example 2 Storage at 2° C.-8° C. Reduces the Rate of Growth ofImpurities in PROCYSBI®

Samples of PROCYSBI® were tested for chemical stability after storageunder conditions of either 2° C.-8° C. (refrigerated)/ambient humidity,or 25° C./60% RH (room temperature), for up to 24 months. Samples wereplaced in controlled environmental chambers, which were maintained atthe described temperature (±2° C.) and relative humidity (±5%). At eachscheduled time point, samples were removed from each storage conditionwithin 1 week of the scheduled date, and held at ambient conditionsuntil analyzed. Amounts of cysteamine bitartrate and the 12 impuritiesthat may grow on stability at 25° C./60% RH (cystamine, Peak A, Peak B,Peak C, 2-hydroxythiomorpholine, Peak D, cystamine tartrate amide,2-hydroxymethylthiazolidine, Peak 5, Peak 6, Peak J, and Peak K), aswell as total related substances (TRS), were measured by HPLC (gradientpump; waters column; dimensions: 150-mm×4.6-mm (i.d.); Xbridge C₁₈packing, 3.5-μm particle size; detection: UV @ 210 nm; Injection Volume:10 μL for assay, 100 μL for related substance (RS) testing; flow rate:1.0 mL/min; column temperature: 40° C.; autosampler temperature: 4° C.;run time 40 minutes). The mobile phases were 23.6 mM 1-octanesulfonicacid sodium and 29.0 mM sodium phosphate pH 2.6/ACN/MeOH 85/3/12 (v/v/v)(mobile phase A) and 0.20 M 1-octanesulfonic acid sodium and 0.10 Msodium phosphate pH 2.6/ACN/MeOH 10/18/72 (v/v/v) (mobile phase B).Elution was performed using a gradient (linear) under the followingparameters:

Time (min.) % A % B 0.0 100 0 2.0 100 0 20.0 60 40 25.0 60 40 25.1 100 040.0 100 0

The limit of quantitation (LOQ) was 0.05%. For graphical purposes,values at or below LOQ are presented at 0.05%. Data for the amount ofcysteamine bitartrate in the samples are calculated relative to thelabel claim of 25 mg or 75 mg (“Assay”). Acid dissolution and bufferdissolution tests were conducted in accordance with the dissolutionmethod of USP 36-NF 31 section <711>. In brief, in the acid dissolutiontest, the composition was mixed with 0.1M HCl for 2 hours, at which timea sample of the fluid was taken and assayed for the amount of cysteaminebitartrate present. In the buffer dissolution test, the composition wasimmediately transferred from the acid solution to a pH 6.8 buffersolution, and the liquid was sampled after either 20 minutes or 30minutes. Graphical representation (e.g., the Y-axes) of acid dissolutionand buffer dissolution data are shown relative to the label claim of 25mg or 75 mg. Graphical (e.g., the Y-axes) and tabular representation ofimpurities data show the amount of a given chemical impurity relative tothe amount of cysteamine bitartrate measured in that chromatogram (i.e.,the ratio of the amount of the impurity to the amount of cysteaminebitartrate, expressed as a percent). Graphical and tabularrepresentations of time in months were also rounded to the nearestinteger.

FIGS. 1A-1G show long-term stability data for two lots of 25 mgPROCYSBI® (Lots A and B) stored at 2° C.-8° C./ambient RH for 24 monthsor at 25° C./60% RH for 12 months. Samples stored at 2° C.-8° C. weretested after storage for 3, 6, 9, 12, 15, 18, 21, and 24 months. Samplesstored at 25° C. were tested after storage for 0, 3, 6, 9, 12, 15, 18,21, and 24 months. At each time point, three 60-count bottles were takenfrom storage and analyzed as described above. FIG. 1A shows the amountof cysteamine bitartrate relative to the label claim of 25 mg (“Assay”)(expressed as a percent), while FIGS. 1B and 1C, respectively, show acidstage dissolution and buffer stage dissolution relative to the labelclaim of 25 mg (expressed as a percent). FIGS. 1D-1G show the amount ofthe impurities cystamine, 2-hydroxythiomorpholine, cystamine tartrateamide, and Peak 6 after storage at 2° C.-8° C./ambient RH for 24 monthsor at 25° C./60% RH for 12 months. At 25° C./60% RH, each of theimpurities increased through 12-month storage. In sharp contrast, thelevels of cystamine, 2-hydroxythiomorpholine, cystamine tartrate amide,and Peak 6 exhibited little to no increases through 24 months at 2°C.-8° C. storage.

Tables 2 and 3 compare 2° C.-8° C. drug product storage with storage at25° C./60% RH for lots A and B, respectively, for the 12 impurities thatmay grow on stability (cystamine, Peak A, Peak B, Peak C,2-hydroxythiomorpholine, Peak D, cystamine tartrate amide,2-hydroxymethylthiazolidine, Peak 5, Peak 6, Peak J, and Peak K), aswell as total related substances (TRS). Tables 2 and 3 show that thelevels of each of these impurities at 2° C.-8° C. exhibited little to nogrowth through 24 months. Tables 2 and 3 also show that the impuritylevels after 18 months storage at 2° C.-8° C. were all lower than orequal to levels after only 12 months storage of 25° C./60% RH. As Tables2 and 3 illustrate, 2° C.-8° C. storage of DP markedly reduces the rateof growth impurities relative to storage at 25° C./60% RH.

TABLE 2 Comparison of Impurities Levels for Lot A After Storage at 2°C.-8° C. and at 25° C./60% RH for Impurities That May Increase o9nStability Impurity Level for Lot A % 2° C.-8° C. 25° C./60% RH ImpurityInitial 18 M 24 M 12 M Cystamine 3.3 3.1 3.2 3.8 Peak A ND ND ND 0.10Peak B  0.05  0.05  0.07 0.06 Peak C ND ND <0.05 0.052-hydroxythiomorpholine  0.47  0.35  0.31 0.62 Peak D ND ND ND 0.07Cystamine Tartrate Amide  0.07  0.09  0.10 0.682-hydroxymethylthiazolidine <0.05 <0.05 ND 0.08 Peak 5 <0.05 <0.05 <0.050.11 Peak 6 <0.05 <0.05 <0.05 0.14 Peak J ND ND ND 0.06 Peak K ND ND ND0.07 TRS 4.4 4.1 4.1 6.2 ND = not detected <0.05 = detected but belowLOQ (0.05%)

TABLE 3 Comparison of Impurities Levels for Lot B After Storage at 2°C.-8° C. and at 25° C./60% RH for impurities That May Increase onStability Impurity Level for Lot B % 2° C.-8° C. 25° C./60% RH ImpurityInitial 18 M 24 M 12 M Cystamine 3.2 2.8 2.9 3.3 Peak A ND ND ND 0.10Peak B <0.05  0.06  0.08 0.06 Peak C ND ND ND <0.052-hydroxythiomorpholine  0.45  0.36  0.30 0.57 Peak D ND ND <0.05 0.06Cystamine Tartrate Amide  0.07  0.08  0.09 0.632-hydroxymethylthiazolidine <0.05 <0.05 ND 0.07 Peak 5 <0.05 <0.05 <0.050.11 Peak 6 <0.05 <0.05 <0.05 0.12 Peak J ND ND <0.05 <0.05 Peak K ND NDND <0.05 TRS 4.2 3.8 4.0 5.5 ND = not detected <0.05 = detected butbelow LOQ (0.05%)

FIGS. 2A-2D show the relative amounts of impurities for the two lots asabove, with the addition of stability data for 4 lots (a third lot of 25mg PROCYSBI®, Lot C, and three 75 mg lots, Lots D, E, and F) stored at2° C.-8° C. for 3 months (i.e., 3 lots of the 25 mg strength and 3 lotsof the 75 mg strength). After three months of storage, three 60-countbottles of 25 mg strength PROCYSBI® and two 250-count bottles of 75 mgstrength PROCYSBI® were taken from storage and analyzed as describedabove. FIGS. 2A-2D show that impurity levels measured through threemonths in the additional samples are consistent with the trends observedin the 24-month data shown in FIGS. 1D-1G and Tables 2 and 3. Stabilitydata for the 75 mg strength lots stored at 2° C.-8° C. are expected totrend with data from the 25 mg strength lots at 2° C.-8° C. because (i)the enteric coated beads are the same (e.g., same API, formulation,components ratios), (ii) the manufacturing process is the same; in fact,beads for 25 mg strength capsules are also encapsulated at the 75 mgstrength (i.e., split batches), (iii) the packaging andproduct-contacting materials are the same, and (iv) the degradationproducts and degradation mechanisms are the same for the two dosagestrengths.

Taken together, these results show that the impurities growth ratesduring storage of PROCYSBI® at 2° C.-8° C. are dramatically lower thanthose observed at 25° C./60% RH. In fact, little to no growth ofimpurities was observed through 24-month storage at 2° C.-8° C. At thesame time, there was no degradation in product performance as measuredby Assay, Acid Stage Dissolution, or Buffer Stage Dissolution afterstorage at 2° C.-8° C. Thus, the lower storage temperature provides aclear improvement in product quality.

Example 3 Predicted Shelf-Life Values for PROCYSBI® when Stored at 2°C.-8° C.

Predicted shelf-life values for PROCYSBI® for storage conditions of 2°C.-8° C. were calculated using the software package SLIMStat® for the 24months of data obtained for two lots of 25 mg PROCYSBI® (Lots A and B)(see Example 2). Table 4 shows SLIMStat® predictions for Lots A and Bindividually. Numeric predictions for several impurities were notavailable due to limited data points with values about LOQ (0.05%) over24 months (i.e., the confidence interval did not exceed the productstability specification); these results are represented in Table 4 bythe start symbol (*).

The shortest predicted shelf-life for 25 mg PROCYSBI® stored at 2° C.-8°C. was 79 months, based on Peak B for Lot B. The longest predictedshelf-life was 2,608 months (i.e., 217 years), based upon the2-hydroxythiomorpholine results for Lot B. The shelf-life predictionsbased upon Assay, and Acid Stage Dissolution and Buffer StageDissolution varied from 39 months up to 493 months. Notably, allSLIMStat® shelf-life predictions for 2° C.-8° C. storage were far beyondthe currently approved expiry of 18 months at room temperature, againillustrating the marked decrease in DP degradation with 2° C.-8° C.storage.

TABLE 4 Predicted Shelf-Life at 2° C.-8° C. Storage Predicted Shelf-Lifeat 2° C.-8° C. (M) Test Lot A Lot B Cystamine * 357 Peak A 266 180 PeakB 118  79 Peak C * * 2-hydroxythiomorpholine * 2608  Peak D * *Cystamine Tartrate Amide 584 941 2-hydroxymethylthiazolidine * * Peak5 * * Peak 6 * * Peak J * * Peak K 764 1018  Total Related Substances *439 Assay^(a)  48  39 Acid Stage Dissolution 493 * Buffer StageDissolution^(b)  69  69 * = Confidence interval of SLIMStat ® predictiondoes not exceed Specification. ^(a)Based upon a lower Assayspecification of 90.0%. ^(b)Based upon a minimum dissolutionspecification of 80% within 20 minutes.

Example 4 Impurity Levels in PROCYSBI® Remain Low after Storage at 2°C.-8° C. for 15 Months Followed by Storage for 3 Months at 25° C.

Samples of PROCYSBI® were tested for chemical stability after storage ata temperature of 2° C.-8° C. for up to 15 months followed by excursionsof up to 3 months at 25° C./60% RH, 30° C./65% RH, or 30° C./75% RH (18months of total storage time). Samples were transferred from onecondition to another on the same day. The methods used for storing andtesting the samples were the same as those described above in Example 2.

FIGS. 3A-3H show stability data for two lots of 25 mg PROCYSBI® (Lots Aand B) at 2-8° C. for 15 months followed by excursions of up 3 months at25° C./60% RH, 30° C./65% RH, or 30° C./75% RH. Levels of the impuritiescystamine (FIGS. 3A and 3B), 2-hydroxythiomorpholine (FIGS. 3C and 3D),cystamine tartrate amide (FIGS. 3E and 3F), and Peak 6 (FIGS. 3G and 3H)are shown as amount of the chemical impurity relative to the amount ofcysteamine bitartrate measured in that chromatogram (i.e., the ratio ofthe amount of the impurity to the amount of cysteamine bitartrate,expressed as a percent). The apparent growth rates of the impuritiesduring the 3-month step-up period generally was similar to the growthrates observed for 25 mg PROCYSBI® stored exclusively at 25° C./60% RH.FIG. 4 also shows that the levels of each of the four impurities afterthe 3-month step-ups (18 months total) were lower than their respectivelevels after only 12 months storage exclusively at 25° C./60% RH.

Tables 5 and 6 show the stability date for 25 mg PROCYSBI® Lots A and B,respectively, for the 12 impurities that may increase on stability for(a) 3-month storage at 25° C./60% RH. 300C/65% RH and 30° C./75% RHafter 15 months initial storage 2° C.-8° C., (b) 25° C./60% RH storagefor 12 months, and (c) 2° C.-8° C. storage at both 15 and 18 months. Asshown in Tables 5 and 6, levels of each of the impurities after thesamples were transferred to and stored at higher temperatures were allnearly all lower than or equal to their respective levels after only 12months storage exclusively at 25° C./60% RH.

TABLEs 5 Results for Storage at 25° C./60% RH, 30° C./65% RH, and 30°C./75% RH After Initial Storage at 2° C.-8° C. for Lot A Impurity Levelsfor Lot A (%) 2° C.-8° C. 15 M −> 18 M Step-Ups* 25° C./60% RH TestInitial 15 M 18 M 25° C./60% 30° C./65% 30° C./75% 12 M Cystamine 3.33.2  3.1 3.3  3.2  3.2  3.8 Peak A ND <0.05  ND ND ND ND 0.10 Peak B 0.05 0.06  0.05 0.05 0.06 0.06 0.06 Peak C ND ND ND ND ND ND <0.052-hydroxythiomorpholine  0.47 0.31  0.35 0.42 0.49 0.50 0.62 Peak D NDND ND ND <0.05  <0.05  0.07 Cystamine Tartrate Amide  0.07 0.09  0.090.22 0.42 0.43 0.68 2-hydroxymethylthiazolidine <0.05 ND <0.05 ND ND ND0.08 Peak 5 <0.05 <0.05  <0.05 0.05 0.07 0.08 0.11 Peak 6 <0.05 0.05<0.05 0.07 0.11 0.11 0.14 Peak J ND ND ND ND <0.05  <0.05  0.06 Peak KND ND ND ND ND ND 0.07 TRS 4.4 4.1 4.1 4.5  4.8  4.8  6.2 *15 M storageat 2° C.-8° C. followed by 3 M storage at 25° C./60% RH, 30° C./65% RHor 30° C./75% RH <0.05 = below LOQ ND = not detected

TABLE 6 Results for Storage at 25° C./60% RH, 30° C./65% RH, and 30°C./75% RH After Initial Storage at 2° C.-8° C. for Lot B Impurity Levelsfor Lot B (%) 2° C.-8° C. 15 M −> 18 M Step-Ups* 25° C./60% RH TestInitial 15 M 18 M 25° C./60% 30° C./65% 30° C./75% 12 M Cystamine 3.22.8  2.8 2.8  3.0  2.9  3.3 Peak A ND <0.05  ND ND ND ND 0.10 Peak B<0.05 0.07  0.06 0.10 0.06 0.06 0.06 Peak C ND ND ND <0.05  ND ND <0.052-hydroxythiomorpholine  0.45 0.30  0.36 0.39 0.48 0.47 0.57 Peak D NDND ND <0.05  <0.05  <0.05  0.06 Cystamine Tartrate Amide  0.07 0.08 0.08 0.21 0.38 0.39 0.63 2-hydroxymethylthiazolidine <0.05 ND <0.050.05 ND ND 0.07 Peak 5 <0.05 <0.05  <0.05 0.06 0.07 0.08 0.11 Peak 6<0.05 0.05 <0.05 0.06 0.10 0.10 0.12 Peak J ND ND ND ND <0.05  <0.05 <0.05 Peak K ND ND ND ND ND ND <0.05 TRS 4.2 3.8  3.8 4.1  4.5  4.4  5.5*15 M storage at 2° C.-8° C. followed by 3 M storage at 25° C./60% RH,30° C./65% RH or 30° C./75% RH <0.05 = below LOQ ND = not detected

Taken together. FIGS. 3A-3G and Tables 5 and 6 show that 25 mg PROCYSBI®stored at 2° C.-8° C. for 15 months, followed by storage at 25° C./60%RH for 3 additional months (18 months total time) resulted in lowerlevels for nearly all of the 19 measured impurities relative to 12months storage at 25° C./60% RH. Moreover, samples tested after storagefor 3 months at more stringent conditions (e.g., 30° C./65% RH and 30°C./75% RH) had lower impurity levels than samples stored for 12 monthsat 25° C./60% RH.

Example 5 Predicted Shelf-Life Values for PROCYSBI® when Stored at 2°C.-8° C. Followed by Storage for 4 Months at 25° C.

An extrapolation analysis was performed to evaluate 4-month storage atroom temperature following 2° C.-8° C. long-term storage. Specifically,the duration of 25° C./60% RH storage required for each impurity toincrease from its assumed level at release to its stabilityspecification level as calculated.

Degradation rates for each of the 12 impurities that may increase onstability at 25° C./60% RH were determined from a date set of 10 lots(G, H, I, J, K, L, M, N, O, and P) using SLIMStat® (Table 7).

TABLE 7 Degradation Rates for PROCYSBI ® at 25° C./60% RH DegradationRates (%.month) (by lot) Impurity G H I J K L M N O P Max. Cystamine0.0390 0.0460 0.0296 −0.0167 −0.0067 0.0621 0.0369 0.0927 0.0952 0.07050.0952 Peak A 0.0000 0.0007 0.0006 −0.0024 −0.0026 0.0017 0.0005 0.00280.0031 0.0005 0.0031 Peak B 0.0082 0.0056 0.0041 0.0030 0.0017 0.00230.0029 0.0043 0.0037 0.0050 0.0082 Peak C 0.0005 0.0008 0.0004 0.00000.0000 0.0010 0.0000 0.0042 0.0035 0.0010 0.0042 2-hydroxythiomorpholine0.0226 0.0202 0.0145 0.0217 0.0127 0.0286 0.0182 0.0372 0.0343 0.02880.0372 Peak D 0.0052 0.0053 0.0052 0.0000 0.0000 0.0045 0.0019 0.01330.0105 0.0066 0.0133 Cystamine Tartrate Amide 0.0539 0.0566 0.05420.0543 0.0578 0.0700 0.0505 0.0690 0.0694 0.0487 0.07002-hydroxymethylthiazolidine 0.0087 0.0065 0.0056 0.0007 0.0000 0.00780.0052 0.0114 0.0110 0.0074 0.0114 Peak 5 0.0055 0.0047 0.0046 0.00130.0000 0.0087 0.0065 0.0095 0.0095 0.0060 0.0095 Peak 6 0.0093 0.00680.0068 0.0020 0.0033 0.0106 0.0051 0.0215 0.0181 0.0104 0.0215 Peak J0.0029 0.0023 0.0012 0.0000 0.0000 0.0021 0.0020 0.0098 0.0072 0.00540.0098 Peak K 0.0026 0.0012 0.0002 0.0010 0.0000 0.0005 0.0000 0.00190.0015 0.0000 0.0026 TRS 0.1367 0.1367 0.1527 0.0367 0.0700 0.21900.1638 0.3053 0.2914 0.2373 0.3053

In calculating the degradation rates to be used in the extrapolationanalysis, the largest individual degradation rate for each of the 12impurities was used (“Highest Degradation Rate”).

The level of each impurity at DP lot release was assumed to be close tothe release specification (see ICH Guidance, Evaluation for StabilityData Q1E, 2003). Numerically, close to release specification was takento be 80% of the release specification (e.g., for an impurity with arelease specification of 0.15%, the level of that impurity upon releasewas taken to be 80% of 0.15%, or 0.12%). The release specificationscurrently approved in the US and EU are identical for the 12 impuritiesthat may grow on stability at 25° C./60% RH. The release specificationsand assumed values at release (i.e., “close” to release specifications)for the 12 impurities are shown in Table 8.

The rates of growth of impurities at 2° C.-8° C. were taken to benegligible, based upon the 24-month real-time stability results Example2. That is, the assumed impurity levels at release were taken to be thesame as values after storage at 2° C.-8° C. for up to 24 months.

Using this information, the duration of 25/60% RH storage required foreach impurity to increase from its assumed level at release (i.e., 80%of the release specification to its stability specification level,t_(Step-Up), was calculated with Equation 1 (assuming linear degradationkinetics).

$\begin{matrix}{t_{{Step} - {Up}} = \frac{\left( {{Stability}\mspace{14mu}{Specification}} \right) - \left( {{{Assumed}\mspace{14mu}{Release}\mspace{14mu}{Level}};{``{Close}"}} \right)}{\left( {{Highest}\mspace{14mu}{Degradation}\mspace{14mu}{Rate}} \right)}} & {{Equation}\mspace{14mu} 1}\end{matrix}$

Table 8 shows the calculated t_(Step-Up) values. For each of the 12impurities that may increase on stability, the t_(Step-Up) value isgreater than or equal to 4 months. In summary, the extrapolationanalysis indicates that PROCYSBI® can be stored at 2° C.-8° C. for 0-24months, followed by storage for 4 months at 25° C. without resulting inhigh levels of impurities. Accordingly, patients would be able to storePROCYSBI®, once they receive it, at room temperature conditions for upto four months. Patient storage at refrigerated conditions is alsosupported through a total storage duration of at least 24 months.

TABLE 8 Extrapolation Analysis of 4-Month Step-Ups To 25° C./60% RHStorage Highest Deg Supports Specifications (%) Rate t_(Step-Up) 4 MStep- Impurity Release Close* Stability (%/month) (months) Up to 25° C.Cystamine 4.0 3.2 5.0 0.0952 19 Yes Peak A 0.20 0.16 0.20 0.0031 13 YesPeak B 0.15 0.12 0.15 0.0082 4 Yes Peak C 0.10 0.08 0.10 0.0042 5 Yes2-hydroxythiomorpholine 0.50 0.40 1.0 0.0372 16 Yes Peak D 0.10 0.080.30 0.0133 17 Yes Cystamine Tartrate Amide 0.15 0.12 1.3 0.0700 17 Yes2-hydroxymethylthiazolidine 0.10 0.08 0.3 0.0114 19 Yes Peak 5 0.10 0.080.30 0.0095 23 Yes Peak 6 0.10 0.08 0.30 0.0215 10 Yes Peak J 0.10 0.080.15 0.0098 7 Yes Peak K 0.10 0.08 0.15 0.0026 27 Yes TRS 5.0 4.0 8.00.3053 13 Yes *Close to Release Specification = 80% * (ReleaseSpecification)

The various embodiments described above can be combined to providefurther embodiments. All of the U.S. patents, U.S. patent applicationpublications. U.S. patent applications, foreign patents, foreign patentapplications and non-patent publications referred to in thisspecification and/or listed in the Application Data Sheet areincorporated herein by reference, in their entirety. Aspects of theembodiments can be modified, if necessary to employ concepts of thevarious patents, applications and publications to provide yet furtherembodiments.

These and other changes can be made to the embodiments in light of theabove-detailed description. In general, in the following claims, theterms used should not be construed to limit the claims to the specificembodiments disclosed in the specification and the claims, but should beconstrued to include all possible embodiments along with the full scopeof equivalents to which such claims are entitled. Accordingly, theclaims are not limited by the disclosure.

The invention claimed is:
 1. A method of lowering intracellular cystinelevels in a subject in need thereof, comprising: administering twicedaily to the subject an oral pharmaceutical composition comprisingcysteamine bitartrate, wherein the oral pharmaceutical compositioncontains 2-hydroxythiomorpholine in an amount less than 0.5% relative tothe amount of cysteamine bitartrate, wherein the oral pharmaceuticalcomposition was stored by a manufacturer, a distributor, a pharmacy, ora hospital at a temperature of between about 2° C. and about 8° C. priorto dispensing the oral pharmaceutical composition to the subject, andwherein after the oral pharmaceutical composition is dispensed to thesubject, the oral pharmaceutical composition is stored at a temperatureof between about 20° C. and about 25° C.
 2. The method according toclaim 1, wherein the oral pharmaceutical composition further comprisesan enteric coating wherein the enteric coating comprisespoly(methacrylic acid co-ethyl acrylate) 1:1.
 3. The method according toclaim 1, wherein the oral pharmaceutical composition contains2-hydroxythiomorpholine in an amount less than 0.5% relative to theamount of cysteamine bitartrate when the oral pharmaceutical compositionis stored at a temperature of between about 2° C. and about 8° C. for aperiod of up to 15 months.
 4. The method according to claim 1, whereinthe oral pharmaceutical composition contains 2-hydroxythiomorpholine inan amount less than 0.5% relative to the amount of cysteamine bitartratewhen the oral pharmaceutical composition is stored at a temperature ofbetween about 2° C. and about 8° C. for an initial period of up to 15months, and wherein the oral pharmaceutical composition is subsequentlystored at a temperature of 25° C. and 60% relative humidity for a periodof up to 3 months.
 5. The method according to claim 1, wherein the oralpharmaceutical composition contains 2-hydroxythiomorpholine in an amountless than 0.5% relative to the amount of cysteamine bitartrate when theoral pharmaceutical composition is stored at a temperature of betweenabout 2° C. and about 8° C. for an initial period of up to 15 months,and wherein the oral pharmaceutical composition is subsequently storedat a temperature of 30° C. and 65% relative humidity for a period of upto 3 months.
 6. The method according to claim 1, wherein the oralpharmaceutical composition contains 2-hydroxythiomorpholine in an amountless than 0.5% relative to the amount of cysteamine bitartrate when theoral pharmaceutical composition is stored at a temperature of betweenabout 2° C. and about 8° C. for an initial period of up to 15 months,and wherein the oral pharmaceutical composition is subsequently storedat a temperature of 30° C. and 70% relative humidity for a period of upto 3 months.
 7. A method of lowering intracellular cystine levels in asubject in need thereof, comprising: administering twice daily to thesubject an oral pharmaceutical composition comprising cysteaminebitartrate, wherein the oral pharmaceutical composition containscystamine in an amount less than 4% relative to the amount of cysteaminebitartrate, wherein the oral pharmaceutical composition was stored by amanufacturer, a distributor, a pharmacy, or a hospital at a temperatureof between about 2° C. and about 8° C. prior to dispensing the oralpharmaceutical composition to the subject, and wherein after the oralpharmaceutical composition is dispensed to the subject, the oralpharmaceutical composition is stored at a temperature of between about20° C. and about 25° C.
 8. The method according to claim 7, wherein theoral pharmaceutical composition further comprises an enteric coating,wherein the enteric coating comprises poly(methacrylic acid co-ethylacrylate) 1:1.
 9. The method according to claim 7, wherein the oralpharmaceutical composition contains cystamine in an amount less than 4%relative to the amount of cysteamine bitartrate when the oralpharmaceutical composition is stored at a temperature of between about2° C. and about 8° C. for a period of up to 15 months.
 10. The methodaccording to claim 7, wherein the oral pharmaceutical compositioncontains cystamine in an amount less than 4% relative to the amount ofcysteamine bitartrate when the oral pharmaceutical composition is storedat a temperature of between about 2° C. and about 8° C. for an initialperiod of up to 15 months, and wherein the oral pharmaceuticalcomposition is subsequently stored at a temperature of 25° C. and 60%relative humidity for a period of up to 3 months.
 11. The methodaccording to claim 7, wherein the oral pharmaceutical compositioncontains cystamine in an amount less than 4% relative to the amount ofcysteamine bitartrate when the oral pharmaceutical composition is storedat a temperature of between about 2° C. and about 8° C. for an initialperiod of up to 15 months, and wherein the oral pharmaceuticalcomposition is subsequently stored at a temperature of 30° C. and 65%relative humidity for a period of up to 3 months.
 12. The methodaccording to claim 7, wherein the oral pharmaceutical compositioncontains cystamine in an amount less than 4% relative to the amount ofcysteamine bitartrate when the oral pharmaceutical composition is storedat a temperature of between about 2° C. and about 8° C. for an initialperiod of up to 15 months, and wherein the oral pharmaceuticalcomposition is subsequently stored at a temperature of 30° C. and 70%relative humidity for a period of up to 3 months.
 13. A method oflowering intracellular cystine levels in a subject in need thereof,comprising: administering twice daily to the subject an oralpharmaceutical composition comprising cysteamine bitartrate, wherein thepharmaceutical composition contains cystamine tartrate amide in anamount less than 0.5% relative to the amount of cysteamine bitartrate,wherein the oral pharmaceutical composition was stored by amanufacturer, a distributor, a pharmacy, or a hospital at a temperatureof between about 2° C. and about 8° C. prior to dispensing the oralpharmaceutical composition to the subject, and wherein after the oralpharmaceutical composition is dispensed to the subject, the oralpharmaceutical composition is stored at a temperature of between about20° C. and about 25° C.
 14. The method according to claim 13, whereinthe oral pharmaceutical composition further comprises an entericcoating, wherein the enteric coating comprises poly(methacrylic acidco-ethyl acrylate) 1:1.
 15. The method according to claim 13, whereinthe oral pharmaceutical composition contains cystamine tartrate amide inan amount less than 0.5% relative to the amount of cysteamine bitartratewhen the oral pharmaceutical composition is stored at a temperature ofbetween about 2° C. and about 8° C. for a period of up to 15 months. 16.The method according to claim 13, wherein the oral pharmaceuticalcomposition contains cystamine tartrate amide in an amount less than0.5% relative to the amount of cysteamine bitartrate when the oralpharmaceutical composition is stored at a temperature of between about2° C. and about 8° C. for an initial period of up to 15 months, andwherein the oral pharmaceutical composition is subsequently stored at atemperature of 25° C. and 60% relative humidity for a period of up to 3months.
 17. The method according to claim 13, wherein the oralpharmaceutical composition contains cystamine tartrate amide in anamount less than 0.5% relative to the amount of cysteamine bitartratewhen the oral pharmaceutical composition is stored at a temperature ofbetween about 2° C. and about 8° C. for an initial period of up to 15months, and wherein the oral pharmaceutical composition is subsequentlystored at a temperature of 30° C. and 65% relative humidity for a periodof up to 3 months.
 18. The method according to claim 13, wherein theoral pharmaceutical composition contains cystamine tartrate amide in anamount less than 0.5% relative to the amount of cysteamine bitartratewhen the oral pharmaceutical composition is stored at a temperature ofbetween about 2° C. and about 8° C. for an initial period of up to 15months, and wherein the oral pharmaceutical composition is subsequentlystored at a temperature of 30° C. and 70% relative humidity for a periodof up to 3 months.